CN111939165B

CN111939165B - Application of non-natural ginsenoside 3 beta-O-Glc-DM in preparation of medicine for preventing or treating glioblastoma

Info

Publication number: CN111939165B
Application number: CN201910403991.0A
Authority: CN
Inventors: 杨金玲; 朱平; 李燕; 姜逢霖; 杜倩倩; 唐梅
Original assignee: Institute of Materia Medica of CAMS
Current assignee: Institute of Materia Medica of CAMS
Priority date: 2019-05-15
Filing date: 2019-05-15
Publication date: 2022-09-16
Anticipated expiration: 2039-05-15
Also published as: CN111939165A

Abstract

The invention relates to application of non-natural ginsenoside 3 beta-O-Glc-DM in preparing anti-brain tumor medicines. The 3 beta-O-Glc-DM provided by the invention can effectively treat brain tumor.

Description

Application of non-natural ginsenoside 3 beta-O-Glc-DM in preparation of medicine for preventing or treating glioblastoma

Technical Field

The invention relates to application of non-natural ginsenoside 3 beta-O-D-glucopyranosyl-dammar-24-ene-3 beta, 20S-diol (3 beta-O-D-glucopyranosyl-dammar-24-ene-3 beta, 20S-diol, 3 beta-O-Glc-DM for short) in preparing anti-brain tumor medicines, belonging to the technical field of medicines.

Background

The non-natural ginsenoside 3 beta-O-Glc-DM has been disclosed to be effective in treating lung cancer and rectal cancer (see patent, application No. 201811151561.6).

The World Health Organization (WHO) classifies primary brain tumors into four categories. WHO grades I and II are low grade gliomas, while anaplastic astrocytomas and anaplastic oligodendrogliomas (WHO grade III) and Glioblastoma (GBM) (WHO grade IV) are collectively referred to as malignant gliomas. Brain glioma is the most common intracranial tumor, accounting for about 50% of all primary nervous system tumors, and is the top in both incidence and mortality of brain malignancies. According to the latest statistical data of the national cancer center, 10.6 cases of new cerebroma in China are shown in 2015, the incidence rate is 7.72/10 ten thousands, and the cerebroma is positioned in the first ten malignant tumors in China. Brain glioma has the characteristic of infiltration and growth, and has great damage to nerve tissues. The surgical resection together with radiotherapy and chemotherapy is a basic means for treating brain glioma, but complete surgical resection is difficult, the slow reaction of glioma cells to rays causes poor chemoradiotherapy effect, and chemotherapy drugs cannot penetrate through the blood brain barrier to play a role, so that the current means for effectively treating brain glioma is lacked, and the recurrence rate is extremely high. The mean survival of patients with brain gliomas is short, the median survival of low-grade malignant gliomas and anaplastic gliomas is generally 3-8 years, and the median survival of GBM is 14.6 months. Temozolomide (TMZ) is the only chemotherapeutic drug with definite curative effect for treating glioma, and the clinical single use of TMZ for treating glioma has unsatisfactory treatment effect due to administration dosage, adverse reaction and drug resistance. Therefore, there is a great need to develop a new therapeutic agent with excellent antitumor activity, in particular with tolerable toxic properties against GBM (and other astrocytic brain tumors).

Disclosure of Invention

The invention solves the technical problem of providing the application of the compound I and the pharmaceutically acceptable salt thereof in preparing anti-brain tumor medicines.

In order to solve the technical problem, the invention provides the following technical scheme:

the first aspect of the technical scheme of the invention provides the application of the compound I and the pharmaceutically acceptable salts thereof in preparing the medicines for preventing or treating the brain tumor,

the chemical name of the compound I is 3 beta-O-D-glucopyranosyl-dammar-24-ene-3 beta, 20S-diol (3 beta-O-D-glucopyranosyl-dammar-24-ene-3 beta, 20S-diol, which is abbreviated as 3 beta-O-Glc-DM).

The brain tumor comprises a glioblastoma.

In a first aspect of the technical scheme of the present invention, an application of a pharmaceutical composition in preparing a medicament for preventing or treating brain tumor is provided, wherein the pharmaceutical composition comprises compound I and a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

The dosage forms of the pharmaceutical composition comprise tablets, capsules, powder, granules, dripping pills, pastes and powders.

The compounds, compositions, and methods described herein can be administered to a subject having or diagnosed with a brain tumor. Various means for administering compound I described herein to a subject are known to those of skill in the art. Such methods may include, but are not limited to: oral administration, parenteral administration, intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, airway administration (aerosol), pulmonary administration, dermal administration, topical administration, injection administration or intratumoral administration. Administration may be local or systemic. In some embodiments, the preferred administration is oral administration.

In some embodiments, compound I or a pharmaceutically acceptable salt thereof is administered alone to the patient as the sole active ingredient.

In some embodiments, a second agent and/or treatment may be further administered to the subject before, after, or concurrently with the administration of compound I or a pharmaceutically acceptable salt thereof, e.g., as part of a combination therapy. The second agent and/or treatment may include a chemotherapeutic agent and/or radiation therapy, and/or surgery.

As used herein, "chemotherapeutic agent" refers to a substance that reduces or diminishes the growth, survival and/or metastasis of cancer cells. Chemotherapeutic agents may include toxins, small molecules, and/or polypeptides. Non-limiting examples of second agents and/or treatments may include: radiotherapy; surgery; taxane antineoplastic agents, vinblastine antineoplastic agents, platinum complexes and/or pyrimidine antagonists.

In some embodiments of the invention, taxane antineoplastic agents include, but are not limited to, paclitaxel and docetaxel; vinblastine antineoplastic agents include, but are not limited to, vinblastine, vincristine, vindesine, and vinorelbine; platinum complexes include, but are not limited to, miriplatin, cisplatin, carboplatin, nedaplatin, and oxaliplatin; pyrimidine antagonists include, but are not limited to, cytarabine, ancitabine, capecitabine, gemcitabine, fluorouracil, bifuran fluorouracil, doxifluridine, tegafur, and carmofur.

In addition, the method of treatment may further comprise the use of radiation or radiotherapy. In addition, the method of treatment may further comprise the use of surgical treatment.

In certain embodiments, an effective dose of compound I, or a pharmaceutically acceptable salt thereof, described herein can be administered to a patient at one time. In certain embodiments, an effective dose of compound I or a pharmaceutically acceptable salt thereof can be repeatedly administered to a patient. For systemic administration, a therapeutically effective amount of compound I or a pharmaceutically acceptable salt thereof may be administered to the patient, including from 0.1mg/kg to 50mg/kg, e.g., 0.1mg/kg, 0.5mg/kg, 1.0mg/kg, 2.0mg/kg, 2.5mg/kg, 5mg/kg, 10mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 30mg/kg, 40mg/kg, 50mg/kg or more.

In some embodiments, treatment may be given on a less frequent basis after the initial treatment regimen. For example, after three months of treatment every two weeks, treatment may be repeated once a month to six months or one year or more. Treatment according to the methods described herein can reduce the level of the marker or a symptom of the disorder, e.g., by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more.

The amount of compound I or a pharmaceutically acceptable salt thereof administered may be determined according to the severity of the disease, the response to the disease, any treatment-related toxicities, the age and health of the patient.

The dosage of compound I or a pharmaceutically acceptable salt thereof described herein can be determined by a clinician and adjusted as necessary to accommodate the observed therapeutic effect. With respect to the duration of treatment and the frequency of treatment, the subject is typically monitored by a skilled clinician to determine when the treatment provides a therapeutic effect and to determine whether to increase or decrease the dosage, increase or decrease the frequency of administration, discontinue treatment, resume treatment, or make other changes to the treatment regimen. The dosing schedule may vary from once a week to once a day, depending on a number of clinical factors, such as the sensitivity of the subject to the composition. The desired dose or amount of active effect may be administered at once or divided into sub-doses, e.g., 2-4 sub-doses, and administered over a period of time (e.g., at appropriate intervals throughout the day or other appropriate schedule). In some embodiments, administration may be chronic, e.g., once or multiple times daily over a period of weeks or months. Examples of dosing and/or treatment schedules are once daily, twice daily, three times daily, four times daily or more dosing over a period of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or longer. The composition may be administered over a period of time, for example, over 5 minutes, 10 minutes, 15 minutes, 20 minutes, or 25 minutes.

In some embodiments of the present invention, the pharmaceutical composition is a formulation suitable for oral administration, including tablets, capsules, powders, granules, dripping pills, pastes, powders, and the like, preferably tablets and capsules. Wherein the tablet can be common tablet, dispersible tablet, effervescent tablet, sustained release tablet, controlled release tablet or enteric coated tablet, and the capsule can be common capsule, sustained release capsule, controlled release capsule or enteric coated capsule. The oral preparation can be prepared by a conventional method using a pharmaceutically acceptable carrier well known in the art. Pharmaceutically acceptable carriers include fillers, absorbents, wetting agents, binders, disintegrants, lubricants, and the like. Fillers include starch, lactose, mannitol, microcrystalline cellulose, and the like; the absorbent comprises calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents include water, ethanol, and the like; the binder comprises hypromellose, polyvidone, microcrystalline cellulose, etc.; the disintegrating agent comprises croscarmellose sodium, crospovidone, surfactant, low-substituted hydroxypropyl cellulose, etc.; the lubricant comprises magnesium stearate, pulvis Talci, polyethylene glycol, sodium laurylsulfate, silica gel micropowder, pulvis Talci, etc. The medicinal adjuvants also include colorant, sweetener, etc.

In some embodiments, the pharmaceutical composition is a solid formulation suitable for oral administration. The composition may be in the form of a tablet or capsule. In a particular embodiment, the pharmaceutical composition is a capsule. In a particular embodiment of the invention, the pharmaceutically acceptable carrier of the oral solid formulation comprises mannitol, microcrystalline cellulose, hydroxypropylcellulose, magnesium stearate.

In some embodiments, a pharmaceutical composition formulated in a unit dosage form for the treatment of brain tumors is provided. In some embodiments, the pharmaceutical composition in unit dosage form contains 2 mg to 20mg of compound I or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition in unit dosage form contains 5mg to 20mg of compound I or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition in unit dosage form contains 8 mg to 20mg of compound I or a pharmaceutically acceptable salt thereof, preferably 10mg to 16 mg of compound I or a pharmaceutically acceptable salt thereof, more preferably 10mg to 14 mg of compound I or a pharmaceutically acceptable salt thereof. In a particular embodiment, the pharmaceutical composition in unit dosage form contains 10mg of compound I or a pharmaceutically acceptable salt thereof. In a particular embodiment, the pharmaceutical composition in unit dosage form contains 12 mg of compound I or a pharmaceutically acceptable salt thereof. In a particular embodiment, the pharmaceutical composition in unit dosage form contains 14 mg of compound I or a pharmaceutically acceptable salt thereof. In a particular embodiment, the pharmaceutical composition in unit dosage form contains 16 mg of compound I or a pharmaceutically acceptable salt thereof. For example, with respect to tablets or capsules, a "pharmaceutical composition in unit dose form" means each tablet or each capsule.

By "pharmaceutically acceptable carrier" is meant that it is used to prepare a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable, and includes that it is acceptable for human pharmaceutical use. Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer solutions, solvents and/or dispersion vehicles. The use of such carriers and diluents is well known in the art. Some non-limiting examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, methyl cellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) lubricants, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol (PEG); (12) esters, e.g. ethyl oleateEsters and ethyl laurate; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) ringer's solution; (19) ethanol; (20) a pH buffer solution; (21) polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids; (23) serum components, such as serum albumin, HDL, and LDL; (24) c ₂ -C ₁₂ Alcohols, such as ethanol; and (25) other non-toxic compatible materials used in pharmaceutical formulations. Wetting agents, coloring agents, mold release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preserving agents and antioxidants can also be present in the formulation. Terms such as "excipient," "carrier," "pharmaceutically acceptable carrier," and the like are used interchangeably herein.

"pharmaceutically acceptable salts" include, but are not limited to, acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with organic acids such as acetic acid, trifluoroacetic acid, propionic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-chlorobenzenesulfonic acid, p-toluenesulfonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid and the like.

By "therapeutically effective amount" is meant an amount of a compound that, when administered to a human for the treatment of a disease, is sufficient to effect treatment for the disease.

By "treatment" is meant any administration of a therapeutically effective amount of a compound and includes:

(1) inhibiting the disease (i.e., arresting the further development of the pathology and/or symptomatology) in a human experiencing or exhibiting the pathology or symptomatology of the disease, or

(2) Ameliorating the disease (i.e., reversing the pathology and/or symptomatology) in a human experiencing or exhibiting the pathology or symptomatology of the disease.

"late" includes "locally late".

Advantageous technical effects

Detailed Description

EXAMPLE 13 preparation of beta-O-Glc-DM (Compound I)

3 beta-O-Glc-DM was prepared according to the method in CN 103849672A.

Pharmacological experiments:

EXAMPLE 13 in vitro anti-tumor assay of beta-O-Glc-DM

The survival rate of tumor cells is determined by MTT method: iressa (Iressa) (purity)>98%, synthesized by the institute of medicine, academy of Chinese medicine) as a positive control. The tumor cell lines include human glioblastoma cell line U87-MG, mouse glioblastoma cell line G422 and GL 261. Digesting the cells in logarithmic phase with pancreatin, preparing single cell suspension with a certain concentration by using a complete culture medium, inoculating the single cell suspension into a 96-well plate according to the difference of cell growth speed according to 3000 cells per well, and adding 100 mu L of cell suspension into each well. Adding 1 × 10 in the next day ^-5 ～5×10 ^-4 Different concentrations of 3 β -O-Glc-DM (in DMSO) or DMSO (solvent control) in mol/L, 100 μ L per well (DMSO final concentration)<0.1%) with three parallel wells per group. At 37 ℃ with 5% CO ₂ After further incubation for 120h the supernatant was discarded and 50. mu.L of freshly prepared serum-free medium containing 2.0mg/mL MTT (Sigma Chemical) was added to each well. Continuously culturing for 4h, removing supernatant, adding 150 μ L DMSO into each well to dissolve MTT formazan precipitate, shaking with micro oscillator, mixing, measuring Optical Density (OD) with enzyme-labeling instrument (WD-2102A, China) at detection wavelength of 570nm, calculating inhibition rate of drug on tumor cells according to the following formula with tumor cells treated with solvent as control group, and calculating IC according to middle effect equation ₅₀ ：

The experimental results show (see table 1) that 3 β -O-Glc-DM has a certain inhibitory effect on the cell growth of glioblastoma cultured in vitro.

TABLE 13 growth inhibition of beta-O-Glc-DM on in vitro cultured tumor cells

Experimental example 23 evaluation of in vivo antitumor Activity of beta-O-Glc-DM

The mouse G422 ectopic cerebroma model is a KM mouse (male, 6-8 weeks old) purchased from the center of laboratory animals of Chinese drug and biological product institute in Beijing, and is bred in an SPF-level animal room with 5 mice per cage, and bred and managed by professionals. The animal room has sufficient illumination, good ventilation and air conditioning equipment, room temperature of 18-25 ℃ and relative humidity of 50-70%. In the experiment, well-grown tumor tissues are taken, cut into pieces, ground and diluted into tumor cell suspension (5X 10) by using sterile normal saline ⁷ Per ml), 0.2ml of tumor fluid was inoculated per mouse axillary back. Animals were randomly assigned, weighed, and dosing was started the following day after inoculation.

TMZ (temozolomide) is used as a control, a mouse G422 brain tumor model and experimental animals are divided into 5 groups, a tumor-bearing solvent control group and TMZ 30.0 mg/kg ^-1 ，3β-O-Glc-DM 5.0mg·kg ^-1 、 10.0mg·kg ^-1 Single component medicine, TMZ 30.0 mg/kg ^-1 With 3 beta-O-Glc-DM 10.0 mg/kg ^-1 Groups of 5 animals were combined. At the end of the experiment, the animals were sacrificed, weighed, tumor tissue stripped and weighed. Calculating tumor inhibition (%) according to weight, tumor weight average + -standard deviation

Indicated, and a t-test between each dosing group and the negative control group was performed.

The results showed that (see Table 2), ginsenoside 3 beta-O-Glc-DM was at 10.0 mg.kg ^-1 When the compound is used alone, the growth inhibition rate of the compound on mouse G422 brain tumor reaches 64.92%, when the compound is used together with TMZ, the inhibition rate can reach 78.71%, and the compound is obviously improved compared with that when the compound is used alone (50.26%).

TABLE 23 growth inhibition of mouse G422 brain tumor by beta-O-Glc-DM

P <0.05, p <0.01, p <0.001, compared to the solvent control group; Δ p <0.05, compared to TMZ group.

The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.

It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims

1. The application of the compound shown in the formula I or the pharmaceutically acceptable salt thereof in preparing the medicine for preventing or treating the glioblastoma,

2. use of a pharmaceutical composition for the manufacture of a medicament for the prevention or treatment of glioblastoma, said pharmaceutical composition comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, according to claim 1, and at least one pharmaceutically acceptable carrier.

3. Use according to claim 2, characterized in that the pharmaceutical composition is in the form of tablets, capsules, granules, pills, pastes, powders.